和营五子方通过促进线粒体自噬的HIF-1α/BNIP3/NIX轴减轻糖尿病视网膜病变。
Heyingwuzi formulation alleviates diabetic retinopathy by promoting mitophagy the HIF-1α/BNIP3/NIX axis.
作者信息
Wu Jia-Jun, Zhang Shu-Yan, Mu Lin, Dong Zhi-Guo, Zhang Yin-Jian
机构信息
Department of Ophthalmology, Longhua Hospital Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Department of Ophthalmology, Eye and ENT Hospital, Fudan University, Shanghai 200031, China.
出版信息
World J Diabetes. 2024 Jun 15;15(6):1317-1339. doi: 10.4239/wjd.v15.i6.1317.
BACKGROUND
Diabetic retinopathy (DR) is the primary cause of visual problems in patients with diabetes. The Heyingwuzi formulation (HYWZF) is effective against DR.
AIM
To determine the HYWZF prevention mechanisms, especially those underlying mitophagy.
METHODS
Human retinal capillary endothelial cells (HRCECs) were treated with high glucose (hg), HYWZF serum, PX-478, or Mdivi-1 . Then, cell counting kit-8, transwell, and tube formation assays were used to evaluate HRCEC proliferation, invasion, and tube formation, respectively. Transmission electron microscopy was used to assess mitochondrial morphology, and Western blotting was used to determine the protein levels. Flow cytometry was used to assess cell apoptosis, reactive oxygen species (ROS) production, and mitochondrial membrane potential. Moreover, C57BL/6 mice were established using streptozotocin and treated with HYWZF for four weeks. Blood glucose levels and body weight were monitored continuously. Changes in retinal characteristics were evaluated using hematoxylin and eosin, tar violet, and periodic acid-Schiff staining. Protein levels in retinal tissues were determined Western blotting, immunohistochemistry, and immunostaining.
RESULTS
HYWZF inhibited excessive ROS production, apoptosis, tube formation, and invasion in hg-induced HRCECs mitochondrial autophagy . It increased the mRNA expression levels of BCL2-interacting protein 3 (), FUN14 domain-containing 1, BNIP3-like (, also known as ), , PTEN-induced kinase 1, and hypoxia-inducible factor (). Moreover, it downregulated the protein levels of vascular endothelial cell growth factor and increased the light chain 3-II/I ratio. However, PX-478 and Mdivi-1 reversed these effects. Additionally, PX-478 and Mdivi-1 rescued the effects of HYWZF by decreasing oxidative stress and apoptosis and increasing mitophagy. HYWZF intervention improved the symptoms of diabetes, tissue damage, number of acellular capillaries, and oxidative stress . Furthermore, experiments confirmed the results of experiments.
CONCLUSION
HYWZF alleviated DR and associated damage by promoting mitophagy the HIF-1α/BNIP3/NIX axis.
背景
糖尿病视网膜病变（DR）是糖尿病患者视力问题的主要原因。和营五子方（HYWZF）对DR有效。
目的
确定HYWZF的预防机制，尤其是线粒体自噬相关机制。
方法
用人视网膜毛细血管内皮细胞（HRCECs）分别用高糖（hg）、HYWZF血清、PX - 478或Mdivi - 1处理。然后，分别用细胞计数试剂盒 - 8、Transwell和管形成实验评估HRCEC的增殖、侵袭和管形成。用透射电子显微镜评估线粒体形态，用蛋白质免疫印迹法测定蛋白质水平。用流式细胞术评估细胞凋亡、活性氧（ROS）产生和线粒体膜电位。此外，用链脲佐菌素建立C57BL/6小鼠模型并用HYWZF治疗四周。连续监测血糖水平和体重。用苏木精 - 伊红、焦油紫和过碘酸 - 希夫染色评估视网膜特征的变化。用蛋白质免疫印迹法、免疫组织化学和免疫染色法测定视网膜组织中的蛋白质水平。
结果
HYWZF抑制hg诱导的HRCECs中线粒体自噬过度的ROS产生、凋亡、管形成和侵袭。它增加了BCL2相互作用蛋白3（ ）、含FUN14结构域1、BNIP3样蛋白（ ，也称为 ）、 、PTEN诱导激酶1和缺氧诱导因子（ ）的mRNA表达水平。此外，它下调了血管内皮生长因子的蛋白质水平并增加了轻链3 - II/I比值。然而，PX - 478和Mdivi - 1逆转了这些作用。此外，PX - 478和Mdivi - 1通过降低氧化应激和凋亡并增加线粒体自噬挽救了HYWZF的作用。HYWZF干预改善了糖尿病症状、组织损伤、无细胞毛细血管数量和氧化应激 。此外， 实验证实了 实验的结果。
结论
HYWZF通过促进线粒体自噬 HIF - 1α/BNIP3/NIX轴减轻DR及相关损伤。
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